Mutant-selective EGFR inhibitors and uses thereof

ABSTRACT

The present invention provides compounds, pharmaceutically acceptable compositions thereof, and methods of using the same.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a continuation application of U.S. patentapplication Ser. No. 14/448,578, filed Jul. 31, 2014 (now U.S. Pat. No.9,409,887), which is a continuation of U.S. patent application Ser. No.13/291,706, filed Nov. 8, 2011 (now U.S. Pat. No. 8,796,255), whichclaims priority to U.S. Provisional Application Nos. 61/412,388, filedNov. 10, 2010, and 61/534,330, filed Sep. 13, 2011, the entirety of eachof which is hereby incorporated by reference.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to compounds useful as mutant-selectiveepidermal growth factor receptor (EGFR) kinase inhibitors. The inventionalso provides pharmaceutically acceptable compositions comprisingcompounds of the present invention and methods of using saidcompositions in the treatment of various disorders.

BACKGROUND OF THE INVENTION

Protein tyrosine kinases are a class of enzymes that catalyze thetransfer of a phosphate group from ATP or GTP to a tyrosine residuelocated on a protein substrate. Receptor tyrosine kinases act totransmit signals from the outside of a cell to the inside by activatingsecondary messaging effectors via a phosphorylation event. A variety ofcellular processes is promoted by these signals, includingproliferation, carbohydrate utilization, protein synthesis,angiogenesis, cell growth, and cell survival.

There is strong precedent for involvement of the EGFR in human cancerbecause over 60% of all solid tumors overexpress at least one of theseproteins or their ligands. Overexpression of EGFR is commonly found inbreast, lung, head and neck, bladder tumors.

Activating mutations in the tyrosine kinase domain of EGFR have beenidentified in patients with non-small cell lung cancer (Lin, N. U.;Winer, E. P., Breast Cancer Res 6: 204-210, 2004). The reversibleinhibitors Tarceva (erlotinib) and Iressa (gefitinib) currently arefirst-line therapy for non-small cell lung cancer patients withactivating mutations. The most common activating mutations are L858R anddelE746-A750.

Additionally, in the majority of patients that relapse, acquired drugresistance, such as by mutation of gatekeeper residue T790M, has beendetected in at least half of such clinically resistant patients.Moreover, T790M may also be pre-existing, there may be an independent,oncogenic role for the T790M mutation. For example, there are patientswith the L858R/T790M mutation who never received gefitinib treatment. Inaddition, germline EGFR T790M mutations are linked with certain familiallung cancers.

Current drugs in development, including second generation covalentinhibitors, such as BIBW2992, HKI-272 and PF-0299804, are effectiveagainst the T790M resistance mutation but exhibit dose-limitingtoxicities due to concurrent inhibition of WT EGFR. Accordingly, thereremains a need to find mutant-selective EGFR kinase inhibitors useful astherapeutic agents.

SUMMARY OF THE INVENTION

It has now been found that compounds of this invention, andpharmaceutically acceptable compositions thereof, are effective asmutant-selective EGFR kinase inhibitors. Such compounds include thosedescribed in detail herein.

Compounds of the present invention, and pharmaceutically acceptablecompositions thereof, are useful for treating cancers associated withone or more EGFR mutations. Such diseases, disorders, or conditionsinclude those described herein.

Compounds provided by this invention are also useful for the study ofkinases in biological and pathological phenomena; the study ofintracellular signal transduction pathways mediated by such kinases; andthe comparative evaluation of new kinase inhibitors.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

1. General Description of Compounds of the Invention

In certain embodiments, the present invention provides a compound offormula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   R¹ is —CF₃ or —Cl;-   W is —O— or —NH—;-   R² is —R or

-   R³ is —R, —C(O)R, or —C(O)OR; and-   each R is independently C₁₋₄ alkyl or C₁₋₄ fluoroalkyl.

In certain embodiments, the present invention provides a compound offormula II:

or a pharmaceutically acceptable salt thereof, wherein:

-   R³ is —R, —C(O)R, or —C(O)OR; and-   each R is independently C₁₋₄ alkyl or C₁₋₄ fluoroalkyl.

In some embodiments, R¹ is —CF₃. In some embodiments, R¹ is —Cl.

In some embodiments, W is —NH—. In some embodiments, W is —O—.

As defined generally above, R³ is —R or

In some embodiments, R³ is —R. In some embodiments, R³ is C₁₋₄ alkyl. Insome embodiments, R³ is isopropyl. In some embodiments, R³ is

As defined generally above, R³ is —R, —C(O)R, or —C(O)OR. In someembodiments, R³ is —R. In some embodiments, R³ is C₁₋₄ fluoroalkyl. Insome embodiments, R³ is —CH₂CH₂F. In some embodiments, R³ is —C(O)R. Insome embodiments, R³ is acetyl. In some embodiments, R³ is —C(O)OR. Insome embodiments, R³ is tert-butoxycarbonyl.

Exemplary compounds of the present invention are set forth in Table 1,below.

TABLE 1 Exemplary Compounds

I-1

I-2

I-3

I-4

I-5

In certain embodiments, the present invention provides a compound setforth in Table 1, above, or a pharmaceutically acceptable salt thereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge etal., describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein byreference. Pharmaceutically acceptable salts of the compounds of thisinvention include those derived from suitable inorganic and organicacids and bases. Examples of pharmaceutically acceptable, nontoxic acidaddition salts are salts of an amino group formed with inorganic acidssuch as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuricacid and perchloric acid or with organic acids such as acetic acid,oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid ormalonic acid or by using other methods used in the art such as ionexchange. Other pharmaceutically acceptable salts include adipate,alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate,propionate, stearate, succinate, sulfate, tartrate, thiocyanate,p-toluenesulfonate, undecanoate, valerate salts, and the like.

Salts derived from appropriate bases include alkali metal, alkalineearth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. Representative alkali oralkaline earth metal salts include sodium, lithium, potassium, calcium,magnesium, and the like. Further pharmaceutically acceptable saltsinclude, when appropriate, nontoxic ammonium, quaternary ammonium, andamine cations formed using counterions such as halide, hydroxide,carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and arylsulfonate.

2. Description of Exemplary Embodiments

As described in detail herein, infra, provided compounds are selectiveinhibitors of at least one mutation of EGFR. It has been surprisinglyfound that provided compounds are selective inhibitors of at least onemutation of EGFR as compared to wild-type (“WT”) EGFR. In certainembodiments, an at least one mutation of EGFR is T790M. In certainembodiments, the at least one mutation of EGFR is a deletion mutation.In some embodiments, the at least one mutation of EGFR is an activatingmutation. In certain embodiments, a provided compound selectivelyinhibits at least one deletion/resistant mutation and at least oneactivating mutation as compared to WT EGFR. In some embodiments, aprovided compound selectively inhibits at least one deletion and/or atleast one point mutation, and is sparing as to WT EGFR inhibition.

A mutation of EGFR can be selected from T790M (resistant or oncogenic),L858R (activating), delE746-A750 (activating), G719S (activating), or acombination thereof.

As used herein, the term “selectively inhibits,” as used in comparisonto inhibition of WT EGFR, means that a provided compound inhibits atleast one mutation of EGFR (i.e., at least one deletion mutation, atleast one activating mutation, or a combination of at least one deletionmutation and at least one activating mutation) in at least one assaydescribed herein (e.g., biochemical or cellular). In some embodiments,the term “selectively inhibits,” as used in comparison to WT EGFRinhibition means that a provided compound is at least 50 times morepotent, at least 45 times, at least 40, at least 35, at least 30, atleast 25, or at least 20 times more potent as an inhibitor of at leastone mutation of EGFR, as defined and described herein, as compared to WTEGFR.

As used herein, the term “sparing as to WT EGFR” means that a selectiveinhibitor of at least one mutation of EGFR, as defined and describedabove and herein, inhibits EGFR at the upper limit of detection of atleast one assay as described herein (e.g., biochemical or cellular asdescribed in detail in Examples 6-8). In some embodiments, the term“sparing as to WT EGFR” means that a provided compound inhibits WT EGFRwith an IC₅₀ of at least 10 μM, at least 9 μM, at least 8 μM, at least 7μM, at least 6 μM, at least 5 μM, at least 3 μM, at least 2 μM, or atleast 1 μM.

In certain embodiments, a provided compound selectively inhibits (a) atleast one activating mutation; and (b) T790M; and (c) is sparing as toWT. In some embodiments, an at least one activating mutation is adeletion mutation. In some embodiments, an at least one activatingmutation is a point mutation. In some embodiments, an activatingmutation is L858R. In some embodiments, an activating mutation is G719S.In some embodiments, an activating mutation is delE746-A750.

In some embodiments, the at least one mutation of EGFR is L858R and/orT790M.

Without wishing to be bound by any particular theory, it is believedthat administration of a provided compound to a patient having at leastone activating mutation may preempt formation of the T790M resistancemutation. Thus, in certain embodiments, the present invention provides amethod for inhibiting an activating mutation in a patient comprisingadministering to the patient a provided compound or composition thereof,as described herein.

One of ordinary skill in the art will appreciate that certain patientshave an oncogenic form of the T790M mutation, i.e., the T790M mutationis present prior to administration to the patient any EGFR inhibitor andis therefore oncogenic. Accordingly, in some embodiments, the presentinvention provides a method for inhibiting oncogenic T790M in a patientcomprising administering to the patient a provided compound orcomposition thereof, as described herein.

Tarceva (erlotinib) and Iressa (gefitinib) are first-line therapies forpatients with activating mutations but exhibit dose-limiting toxicitiesdue to concurrent inhibition of WT EGFR. In addition, drugs currently indevelopment, including second generation covalent inhibitors, such asBIBW2992, HKI-272 and PF-0299804, are effective against the T790Mresistance mutation but exhibit dose-limiting toxicities due toconcurrent inhibition of WT EGFR.

It has been surprisingly found that provided compounds selectivelyinhibit each of the EGFR activating and deletion mutations. Moreover,provided compounds are sparing for WT EGFR and associated dose-limitingtoxicities. This stands in contrast to other known EGFR inhibitors(e.g., BIBW2992 and HKI-272) which are only somewhat effective againstmutants but retain activity against WT EGFR and are therefore limited bytoxicities associated with inhibition of WT EGFR.

In some embodiments, a provided compound is at least 50, at least 45times, at least 40, at least 35, at least 30, at least 25, or at least20 times more potent for at least one mutation of EGFR as compared to WTEGFR, as determined by the biochemical assay described in detail inExample 6, infra. In certain embodiments, a provided compound is atleast 20, at least 15, or at least 10 times more potent for at least onemutation of EGFR as compared to WT EGFR, as determined by the cellularassay described in detail in Example 8, infra.

In some embodiments, a provided compound is at least 50, at least 45times, at least 40, at least 35, at least 30, at least 25, or at least20 times more potent for at least one deletion mutation of EGFR ascompared to WT EGFR, as determined by the biochemical assay described indetail in Example 6, infra. In certain embodiments, a provided compoundis at least 20, at least 15, or at least 10 times more potent for atleast one deletion mutation of EGFR as compared to WT EGFR, asdetermined by the cellular assay described in detail in Example 8,infra.

In some embodiments, a provided compound is at least 50, at least 45times, at least 40, at least 35, at least 30, at least 25, or at least20 times more potent for L858R and/or T790M mutation of EGFR as comparedto WT EGFR, as determined by the biochemical assay described in detailin Example 6, infra. In certain embodiments, a provided compound is atleast 20, at least 15, or at least 10 times more potent for L858R and/orT790M mutation of EGFR as compared to WT EGFR, as determined by thecellular assay described in detail in Example 8, infra.

In some embodiments, a provided compound is at least 20, at least 15, orat least 10 times more potent for double mutant in H1975 cells, ascompared to WT EGFR, in the signaling assay described in detail inExample 7.

In certain embodiments, a provided compound inhibits at least onemutation of EGFR selectively as compared to WT EGFR and as compared toother protein kinases (e.g., ErbB2, ErbB4, a TEC-kinase, and/or JAK3).It will be appreciated that the acrylamide moiety, depicted in formulaI, is a warhead group for covalently binding to a key cysteine residuein the binding domain of at least one mutation of EGFR selectively ascompared to WT EGFR and other protein kinases. Protein kinases having acysteine residue in the binding domain are known to one of ordinaryskill in the art. Such protein kinases having a cysteine residue in thebinding domain include the TEC-family of protein kinases (including TEC,BTK, ITK, BMX, JAK3, and RLK). In certain embodiments, the cysteineresidue is conserved across a protein kinase sub-family, such as ErbB1(commonly referred to as EGFR), ErbB2, and ErbB4.

Without wishing to be bound by any particular theory, it is believedthat provided compounds irreversibly inhibit (i.e., covalently modify)at least one mutation of EGFR selectively as compared to WT EGFR andother protein kinases. In some embodiments, a provided compoundirreversibly inhibits at least one mutation of EGFR selectively ascompared to at least one protein kinase selected from ErbB1, ErbB2,ErbB4, TEC, BTK, ITK, BMX, JAK3, or RLK.

Notwithstanding, in certain embodiments, provided compounds do notappreciably inhibit, either reversibly or irreversibly, other proteinkinases. In some embodiments, a provided compound is selective forinhibiting at least one mutatnt of EGFR as compared to off-targetprotein kinases thereby avoiding effects and toxicities associated withinhibition thereof.

3. Uses, Formulation and Administration

Pharmaceutically Acceptable Compositions

According to another embodiment, the invention provides a compositioncomprising a compound of this invention, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier,adjuvant, or vehicle. An amount of compound in a composition of thisinvention is such that is effective to measurably inhibit a proteinkinase, particularly to inhibit at least one mutant of EGFR selectivelyas compared to WT EGFR, in a biological sample or in a patient. Incertain embodiments, an at least one mutant of EGFR is T790M. In certainembodiments, the at least one mutant of EGFR is a deletion mutation ofEGFR. In some embodiments, the at least one mutation of EGFR is L858Rand/or T790M.

In certain embodiments, an amount of compound in a provided compositionis such that is effective to measurably inhibit at least one mutation ofEGFR selectively as compared to WT EGFR and other protein kinases (e.g.,ErbB2, ErbB4, a TEC-kinase, and/or JAK3).

In certain embodiments, the amount of compound in a provided compositionis such that is effective to measurably inhibit at least one mutant ofEGFR selectively as compared to WT EGFR, in a biological sample or in apatient. In certain embodiments, a composition of this invention isformulated for administration to a patient in need of such composition.In some embodiments, a composition of this invention is formulated fororal administration to a patient.

In certain embodiments, the amount of compound in a provided compositionis such that is effective to measurably inhibit at least one mutant ofEGFR selectively as compared to WT EGFR and other protein kinases (e.g.,ErbB2, ErbB4, a TEC-kinase, and/or JAK3), in a biological sample or in apatient. In certain embodiments, a composition of this invention isformulated for administration to a patient in need of such composition.In some embodiments, a composition of this invention is formulated fororal administration to a patient.

The term “patient”, as used herein, means an animal, preferably amammal, and most preferably a human.

The term “pharmaceutically acceptable carrier, adjuvant, or vehicle”refers to a nontoxic carrier, adjuvant, or vehicle that does not destroythe pharmacological activity of the compound with which it isformulated. Pharmaceutically acceptable carriers, adjuvants or vehiclesthat may be used in the compositions of this invention include, but arenot limited to, ion exchangers, alumina, aluminum stearate, lecithin,serum proteins, such as human serum albumin, buffer substances such asphosphates, glycine, sorbic acid, potassium sorbate, partial glyceridemixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethylcellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,polyethylene glycol and wool fat.

Compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously. Sterile injectable forms of thecompositions of this invention may be aqueous or oleaginous suspension.These suspensions may be formulated according to techniques known in theart using suitable dispersing or wetting agents and suspending agents.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a nontoxic parenterally acceptable diluent orsolvent, for example as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, or similar dispersing agents that are commonly used inthe formulation of pharmaceutically acceptable dosage forms includingemulsions and suspensions. Other commonly used surfactants, such asTweens, Spans and other emulsifying agents or bioavailability enhancerswhich are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

Pharmaceutically acceptable compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, pharmaceutically acceptable compositions of thisinvention may be administered in the form of suppositories for rectaladministration. These can be prepared by mixing the agent with asuitable non-irritating excipient that is solid at room temperature butliquid at rectal temperature and therefore will melt in the rectum torelease the drug. Such materials include cocoa butter, beeswax andpolyethylene glycols.

Pharmaceutically acceptable compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, provided pharmaceutically acceptablecompositions may be formulated in a suitable ointment containing theactive component suspended or dissolved in one or more carriers.Carriers for topical administration of compounds of this inventioninclude, but are not limited to, mineral oil, liquid petrolatum, whitepetrolatum, propylene glycol, polyoxyethylene, polyoxypropylenecompound, emulsifying wax and water. Alternatively, providedpharmaceutically acceptable compositions can be formulated in a suitablelotion or cream containing the active components suspended or dissolvedin one or more pharmaceutically acceptable carriers. Suitable carriersinclude, but are not limited to, mineral oil, sorbitan monostearate,polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol,benzyl alcohol and water.

For ophthalmic use, provided pharmaceutically acceptable compositionsmay be formulated as micronized suspensions in isotonic, pH adjustedsterile saline, or, preferably, as solutions in isotonic, pH adjustedsterile saline, either with or without a preservative such asbenzylalkonium chloride. Alternatively, for ophthalmic uses, thepharmaceutically acceptable compositions may be formulated in anointment such as petrolatum.

Pharmaceutically acceptable compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

In certain embodiments, pharmaceutically acceptable compositions of thisinvention are formulated for oral administration.

The amount of compounds of the present invention that may be combinedwith the carrier materials to produce a composition in a single dosageform will vary depending upon the host treated, the particular mode ofadministration. Preferably, provided compositions should be formulatedso that a dosage of between 0.01-100 mg/kg body weight/day of theinhibitor can be administered to a patient receiving these compositions.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of a compound of the present invention in the composition willalso depend upon the particular compound in the composition.

Uses of Compounds and Pharmaceutically Acceptable Compositions

Compounds and compositions described herein are generally useful for theselective inhibition of at least one mutant of EGFR as compared to WTEGFR. In certain embodiments, an at least one mutant of EGFR is T790M.In certain embodiments, the at least one mutant of EGFR is a deletionmutation of EGFR, an activating mutation of EGFR, or a combinationthereof. In some embodiments, the at least one mutation of EGFR is L858Rand/or T790M.

In certain embodiments, a provided compound selectively inhibits (a) atleast one activating mutation, (b) T790M and (c) is sparing as to WT. Insome embodiments, an at least one activating mutation is a deletionmutation. In some embodiments, an at least one activating mutation is apoint mutation. In some embodiments, an activating mutation isdelE746-A750. In some embodiments, an activating mutation is L858R. Insome embodiments, an activating mutation is G719S.

In some embodiments, the at least one mutation of EGFR is L858R and/orT790M.

The activity of a compound utilized in this invention as a selectiveinhibitor of at least one mutant of EGFR as compared to WT EGFR, may beassayed in vitro, in vivo or in a cell line. In vitro assays includeassays that determine inhibition of the phosphorylation activity and/orthe subsequent functional consequences, or ATPase activity of activatedEGFR (WT or mutant). Alternate in vitro assays quantitate the ability ofthe inhibitor to bind to EGFR (WT or mutant). Inhibitor binding may bemeasured by radiolabeling the inhibitor prior to binding, isolating theinhibitor/EGFR (WT or mutant) complex and determining the amount ofradiolabel bound. Alternatively, inhibitor binding may be determined byrunning a competition experiment where new inhibitors are incubated withEGFR (WT or mutant) bound to known radioligands. Detailed conditions forassaying a compound utilized in this invention as an inhibitor of EGFR(WT or mutant), are set forth in the Examples below.

Protein tyrosine kinases are a class of enzymes that catalyze thetransfer of a phosphate group from ATP or GTP to a tyrosine residuelocated on a protein substrate. Receptor tyrosine kinases act totransmit signals from the outside of a cell to the inside by activatingsecondary messaging effectors via a phosphorylation event. A variety ofcellular processes are promoted by these signals, includingproliferation, carbohydrate utilization, protein synthesis,angiogenesis, cell growth, and cell survival.

As used herein, the terms “treatment,” “treat,” and “treating” refer toreversing, alleviating, delaying the onset of, or inhibiting theprogress of a disease or disorder, or one or more symptoms thereof, asdescribed herein. In some embodiments, treatment may be administeredafter one or more symptoms have developed. In other embodiments,treatment may be administered in the absence of symptoms. For example,treatment may be administered to a susceptible individual prior to theonset of symptoms (e.g., in light of a history of symptoms and/or inlight of genetic or other susceptibility factors). Treatment may also becontinued after symptoms have resolved, for example to prevent or delaytheir recurrence.

Provided compounds are inhibitors of at least one mutant of EGFR and aretherefore useful for treating one or more disorders associated withactivity of one of more EGFR mutants (e.g., a deletion mutation, anactivating mutation, or combination thereof). Thus, in certainembodiments, the present invention provides a method for treating amutant EGFR-mediated disorder comprising the step of administering to apatient in need thereof a compound of the present invention, orpharmaceutically acceptable composition thereof.

As used herein, the term “mutant EGFR-mediated” disorders or conditionsas used herein means any disease or other deleterious condition in whichat least one mutant of EGFR is known to play a role. In certainembodiments, an at least one mutant of EGFR is T790M. In someembodiments, the at least one mutant of EGFR is a deletion mutation. Incertain embodiments, the at least one mutant of EGFR is an activatingmutation. In some embodiments, the at least one mutant of EGFR is L858Rand/or T790M. In certain embodiments, a provided compound selectivelyinhibits (a) at least one activating mutation, (b) T790M, and (c) issparing as to WT. In some embodiments, an at least one activatingmutation is a deletion mutation. In some embodiments, an at least oneactivating mutation is a point mutation. In some embodiments, anactivating mutation is delE746-A750. In some embodiments, an activatingmutation is L858R. In some embodiments, an activating mutation is G719S.

Accordingly, another embodiment of the present invention relates totreating or lessening the severity of one or more diseases in which atleast one mutant of EGFR is known to play a role. Specifically, thepresent invention relates to a method of treating or lessening theseverity of a disease or condition selected from a proliferativedisorder, wherein said method comprises administering to a patient inneed thereof a compound or composition according to the presentinvention.

In some embodiments, the present invention provides a method fortreating or lessening the severity of one or more disorders selectedfrom a cancer. In some embodiments, the cancer is associated with asolid tumor. In certain embodiments, the cancer is breast cancer,glioblastoma, lung cancer, cancer of the head and neck, colorectalcancer, bladder cancer, or non-small cell lung cancer. In someembodiments, the present invention provides a method for treating orlessening the severity of one or more disorders selected from squamouscell carcinoma, salivary gland carcinoma, ovarian carcinoma, orpancreatic cancer.

In certain embodiments, the present invention provides a method fortreating or lessening the severity of neurofibromatosis type I (NF1),neurofibromatosis type II (NF2) Schwann cell neoplasms (e.g. MPNST's),or Schwannomas.

The compounds and compositions, according to the method of the presentinvention, may be administered using any amount and any route ofadministration effective for treating or lessening the severity of acancer. The exact amount required will vary from subject to subject,depending on the species, age, and general condition of the subject, theseverity of the infection, the particular agent, its mode ofadministration, and the like. Compounds of the invention are preferablyformulated in dosage unit form for ease of administration and uniformityof dosage. The expression “dosage unit form” as used herein refers to aphysically discrete unit of agent appropriate for the patient to betreated. It will be understood, however, that the total daily usage ofthe compounds and compositions of the present invention will be decidedby the attending physician within the scope of sound medical judgment.The specific effective dose level for any particular patient or organismwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The term “patient”, as usedherein, means an animal, preferably a mammal, and most preferably ahuman.

Pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, the compounds of the invention may be administeredorally or parenterally at dosage levels of about 0.01 mg/kg to about 50mg/kg or from about 1 mg/kg to about 25 mg/kg, of subject body weightper day, one or more times a day, to obtain the desired therapeuticeffect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

Injectable formulations can be sterilized, for example, by filtrationthrough a bacterial-retaining filter, terminal (heat) sterilization, orsterilization via ionizing radiationor by incorporating sterilizingagents in the form of sterile solid compositions which can be dissolvedor dispersed in sterile water or other sterile injectable medium priorto use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like. In some embodiments,a solid composition is a liquid filled hard gelatin capsule or soliddispersion.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

According to another embodiment, the invention relates to a method ofinhibiting at least one mutant of EGFR (e.g., a deletion mutation, anactivating mutation, a resistant mutation, or combination thereof)activity in a biological sample comprising the step of contacting saidbiological sample with a compound of this invention, or a compositioncomprising said compound. In certain embodiments, the invention relatesto a method of irreversibly inhibiting at least one mutant of EGFR(e.g., a deletion mutation, an activating mutation, or combinationthereof) activity in a biological sample comprising the step ofcontacting said biological sample with a compound of this invention, ora composition comprising said compound.

In certain embodiments, a provided compound selectively inhibits in abiological sample (a) at least one activating mutation, (b) T790M, and(c) is sparing as to WT. In some embodiments, an at least one activatingmutation is a deletion mutation. In some embodiments, an at least oneactivating mutation is a point mutation. In some embodiments, anactivating mutation is delE746-A750. In some embodiments, an activatingmutation is L858R. In some embodiments, an activating mutation is G719S.

The term “biological sample”, as used herein, includes, withoutlimitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; and blood, saliva, urine,feces, semen, tears, or other body fluids or extracts thereof.

Inhibition of at least one mutant of EGFR (e.g., a deletion mutation, anactivating mutation, a resistant mutation, or combination thereof)activity in a biological sample is useful for a variety of purposes thatare known to one of skill in the art. Examples of such purposes include,but are not limited to, blood transfusion, organ transplantation,biological specimen storage, and biological assays.

Another embodiment of the present invention relates to a method ofinhibiting at least one mutant of EGFR (e.g., a deletion mutation, anactivating mutation, a resistant mutation, or combination thereof)activity in a patient comprising the step of administering to saidpatient a compound of the present invention, or a composition comprisingsaid compound. In certain embodiments, the present invention provides amethod for inhibiting (a) at least one activating mutation, (b) T790M ina patient, and (c) is sparing as to WT, wherein said method comprisesadministering to the patient a provided compound, or compositionthereof. In some embodiments, an at least one activating mutation is adeletion mutation. In some embodiments, the present invention provides amethod for inhibiting at least one mutant of EGFR in a patient, whereinan at least one activating mutation is a point mutation. In someembodiments, the present invention provides a method for inhibiting atleast one mutant of EGFR in a patient, wherein an activating mutation isdelE746-A750. In some embodiments, the present invention provides amethod for inhibiting at least one mutant of EGFR in a patient, whereinan activating mutation is L858R. In some embodiments, the presentinvention provides a method for inhibiting at least one mutant of EGFRin a patient, wherein an activating mutation is G719S.

According to another embodiment, the invention relates to a method ofinhibiting at least one mutant of EGFR (e.g., a deletion mutation, anactivating mutation, a resistant mutation, or combination thereof)activity in a patient comprising the step of administering to saidpatient a compound of the present invention, or a composition comprisingsaid compound. According to certain embodiments, the invention relatesto a method of irreversibly inhibiting at least one mutant of EGFRactivity (e.g., a deletion mutation, an activating mutation, a resistantmutation, or combination thereof) in a patient comprising the step ofadministering to said patient a compound of the present invention, or acomposition comprising said compound. In certain embodiments, thepresent invention provides a method for irreversibly inhibiting (a) atleast one activating mutation and (b) T790M in a patient, and (c) issparing as to WT, wherein said method comprises administering to thepatient a provided compound, or composition thereof. In someembodiments, an at least one activating mutation is a deletion mutation.In some embodiments, an at least one activating mutation is a pointmutation. In some embodiments, the present invention provides a methodfor irreversibly inhibiting at least one mutant of EGFR in a patient,wherein an activating mutation is delE746-A750. In some embodiments, thepresent invention provides a method for irreversibly inhibiting at leastone mutant of EGFR in a patient, wherein an activating mutation isL858R. In some embodiments, the the present invention provides a methodfor irreversibly inhibiting at least one mutant of EGFR in a patient,wherein an activating mutation is G719S.

In other embodiments, the present invention provides a method fortreating a disorder mediated by one or more of at least one mutant ofEGFR (e.g., a deletion mutation, an activating mutation, a resistantmutation, or combination thereof) in a patient in need thereof,comprising the step of administering to said patient a compoundaccording to the present invention or pharmaceutically acceptablecomposition thereof. Such disorders are described in detail herein.

Depending upon the particular condition, or disease, to be treated,additional therapeutic agents, which are normally administered to treatthat condition, may also be present in the compositions of thisinvention. As used herein, additional therapeutic agents that arenormally administered to treat a particular disease, or condition, areknown as “appropriate for the disease, or condition, being treated.”

For example, compounds of the present invention, or a pharmaceuticallyacceptable composition thereof, are administered in combination withchemotherapeutic agents to treat proliferative diseases and cancer.Examples of known chemotherapeutic agents include, but are not limitedto, Adriamycin, dexamethasone, vincristine, cyclophosphamide,fluorouracil, topotecan, taxol, interferons, platinum derivatives,taxane (e.g., paclitaxel), vinca alkaloids (e.g., vinblastine),anthracyclines (e.g., doxorubicin), epipodophyllotoxins (e.g.,etoposide), cisplatin, an mTOR inhibitor (e.g., a rapamycin),methotrexate, actinomycin D, dolastatin 10, colchicine, emetine,trimetrexate, metoprine, cyclosporine, daunorubicin, teniposide,amphotericin, alkylating agents (e.g., chlorambucil), 5-fluorouracil,campthothecin, cisplatin, metronidazole, and Gleevec™, among others. Inother embodiments, a compound of the present invention is administeredin combination with a biologic agent, such as Avastin or VECTIBIX.

In certain embodiments, compounds of the present invention, or apharmaceutically acceptable composition thereof, are administered incombination with an antiproliferative or chemotherapeutic agent selectedfrom any one or more of abarelix, aldesleukin, alemtuzumab,alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenictrioxide, asparaginase, azacitidine, BCG Live, bevacuzimab,fluorouracil, bexarotene, bleomycin, bortezomib, busulfan, calusterone,capecitabine, camptothecin, carboplatin, carmustine, celecoxib,cetuximab, chlorambucil, cladribine, clofarabine, cyclophosphamide,cytarabine, dactinomycin, darbepoetin alfa, daunorubicin, denileukin,dexrazoxane, docetaxel, doxorubicin (neutral), doxorubicinhydrochloride, dromostanolone propionate, epirubicin, epoetin alfa,erlotinib, estramustine, etoposide phosphate, etoposide, exemestane,filgrastim, floxuridine fludarabine, fulvestrant, gefitinib,gemcitabine, gemtuzumab, goserelin acetate, histrelin acetate,hydroxyurea, ibritumomab, idarubicin, ifosfamide, imatinib mesylate,interferon alfa-2a, interferon alfa-2b, irinotecan, lenalidomide,letrozole, leucovorin, leuprolide acetate, levamisole, lomustine,megestrol acetate, melphalan, mercaptopurine, 6-MP, mesna, methotrexate,methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone,nelarabine, nofetumomab, oprelvekin, oxaliplatin, paclitaxel,palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim,pemetrexed disodium, pentostatin, pipobroman, plicamycin, porfimersodium, procarbazine, quinacrine, rasburicase, rituximab, sargramostim,sorafenib, streptozocin, sunitinib maleate, talc, tamoxifen,temozolomide, teniposide, VM-26, testolactone, thioguanine, 6-TG,thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin,ATRA, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine,zoledronate, or zoledronic acid.

Other examples of agents the inhibitors of this invention may also becombined with include, without limitation: treatments for Alzheimer'sDisease such as donepezil hydrochloride (Aricept®) and rivastigmine(Exelon®); treatments for Parkinson's Disease such as L-DOPA/carbidopa,entacapone, ropinrole, pramipexole, bromocriptine, pergolide,trihexephendyl, and amantadine; agents for treating Multiple Sclerosis(MS) such as beta interferon (e.g., Avonex® and Rebif®), glatirameracetate (Copaxone®), and mitoxantrone; treatments for asthma such asalbuterol and montelukast (Singulair®); agents for treatingschizophrenia such as zyprexa, risperdal, seroquel, and haloperidol;anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1 RA,azathioprine, cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophophamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; and agents for treatingimmunodeficiency disorders such as gamma globulin.

In certain embodiments, compounds of the present invention, or apharmaceutically acceptable composition thereof, are administered incombination with a monoclonal antibody or an siRNA therapeutic.

Those additional agents may be administered separately from an inventivecompound-containing composition, as part of a multiple dosage regimen.Alternatively, those agents may be part of a single dosage form, mixedtogether with a compound of this invention in a single composition. Ifadministered as part of a multiple dosage regime, the two active agentsmay be submitted simultaneously, sequentially or within a period of timefrom one another normally within five hours from one another.

As used herein, the term “combination,” “combined,” and related termsrefers to the simultaneous or sequential administration of therapeuticagents in accordance with this invention. For example, a compound of thepresent invention may be administered with another therapeutic agentsimultaneously or sequentially in separate unit dosage forms or togetherin a single unit dosage form. Accordingly, the present inventionprovides a single unit dosage form comprising a provided compound, anadditional therapeutic agent, and a pharmaceutically acceptable carrier,adjuvant, or vehicle.

The amount of both, an inventive compound and additional therapeuticagent (in those compositions which comprise an additional therapeuticagent as described above)) that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. Preferably,compositions of this invention should be formulated so that a dosage ofbetween 0.01-100 mg/kg body weight/day of an inventive can beadministered.

In those compositions which comprise an additional therapeutic agent,that additional therapeutic agent and the compound of this invention mayact synergistically. Therefore, the amount of additional therapeuticagent in such compositions will be less than that required in amonotherapy utilizing only that therapeutic agent. In such compositionsa dosage of between 0.01-1,000 μg/kg body weight/day of the additionaltherapeutic agent can be administered.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

The compounds of this invention, or pharmaceutical compositions thereof,may also be incorporated into compositions for coating an implantablemedical device, such as prostheses, artificial valves, vascular grafts,stents and catheters. Vascular stents, for example, have been used toovercome restenosis (re-narrowing of the vessel wall after injury).However, patients using stents or other implantable devices risk clotformation or platelet activation. These unwanted effects may beprevented or mitigated by pre-coating the device with a pharmaceuticallyacceptable composition comprising a kinase inhibitor. Implantabledevices coated with a compound of this invention are another embodimentof the present invention.

EXEMPLIFICATION

As depicted in the Examples below, in certain exemplary embodiments,compounds are prepared according to the following general procedures. Itwill be appreciated that, although the general methods depict thesynthesis of certain compounds of the present invention, the followinggeneral methods, and other methods known to one of ordinary skill in theart, can be applied to all compounds and subclasses and species of eachof these compounds, as described herein.

Compound numbers utilized in the Examples below correspond to compoundnumbers set forth in Table 1, supra.

Provided compounds are prepared according to methods known to one ofordinary skill in the art and include methods described in detail in US20100029610, published Feb. 4, 2010, the entirety of which is herebyincorporated herein by reference.

General Methods for Preparing Certain Aniline Intermediates

Into a 100 mL 3-Neck RBF equipped with reflux condenser were charged4-fluoro-2-methoxy-1-nitrobenzene (7.8 g) and tert-butyl3-aminoazetidine-1-carboxylate (8.63 g), TEA (13.8 g) in dimethylsulfoxide (30 mL). The reaction was heated at 95° C. for 8 hours. Aftercompletion of the reaction, the reaction mixture was poured into water(300 mL). Product was extracted with ethyl acetate (3×100 mL). Organiclayer was washed with brine (2×100 mL) and dried over sodium sulfate.Organic layer was concentrated completely under reduce pressure at 40°C. to give compound 1 of Scheme 1 (14.0 g): Calculated mass forC₁₅H₂₁N₃O₅: 323.2, found: 324.3 (M+H⁺).

To compound 1 of Scheme 1 (22 g) in a 250 mL 3-neck RBF was added TFA(50 mL) at room temperature. The reaction mixture was stirred for 15minutes at room temperature. After completion of the reaction, thereaction mixture was concentrated under reduced pressure at 40° C. togive compound 2 of Scheme 1: Calculated mass for C₁₀H₁₃N₃O₃: 223.1,found: 224.0 (M+H⁺).

To compound 2 of Scheme 1 (10 g) in 250 mL 3-Neck RBF were added CS₂CO₃(28.9 g) and 1,2-bromofluoroethane (3.38 g) in 120 mL of acetonitrile.The reaction was heated at 50° C. for 8 hours. After completion of thereaction, the reaction mixture was poured in water. Product wasextracted in ethyl acetate (3×100 mL). The organic layer was washed withbrine (2×100 mL) and dried over sodium sulfate. The organic layer wasconcentrated completely under reduce pressure at 40° C. to give compound3 of Scheme 1 (5.0 g): Calculated mass for C₁₂H₁₆FN₃O₃: 269.1, found:269.9 (M+H⁺).

A solution of compound 3 of Scheme 1 (1.0 g) and Pd/C (0.20 g) in1,4-dioxane (45 mL) was hydrogenated for 4 hours at room temperature.After completion of the reaction, the reaction mixture was filteredthrough Celite-bed. The Celite bed was washed with 10 mL of 1,4-dioxane.The filtrate was concentrated to provide the intermediate 2 of Scheme 1:Calculated mass for C₁₂H₁₈FN₃O: 239.1, found: 239.9 (M+H⁺).

Intermediate 1 of Scheme 1 was obtained by hydrogenation of compound 1of Scheme 1 using Pd/C in 1,4-dioxane as described above: Calculatedmass for C₁₅H₂₃N₃O₃: 293.2, found: 294.2 (M+H⁺).

Intermediate 3 of Scheme 1 was obtained by acetylation of 2 using aceticanhydride and DMAP, followed by hydrogenation using Pd/C in 1,4-dioxaneas described above: Calculated mass for C₁₂H₁₇N₃O₂: 235.3, found: 236.2(M+H⁺).

Example 1 Compound I-1,N-(3-(2-(4-(1-(2-fluoroethyl)azetidin-3-ylamino)-2-methoxyphenylamino)-5-(trifluoromethyl)pyrimidin-4-ylamino)phenyl)acrylamide

Step 1:

Into a 25 mL 3-neck RBF previously equipped with a magnetic stirrer,thermo pocket and CaCl₂ guard tube, N-Boc-1,3-diaminobenzene (0.96 g)and 1-butanol (9.0 mL) were charged. Reaction mixture was cooled to 0°C. 2,4-Dichloro-5-trifluoromethylpyrimidine (1.0 g) was added dropwiseto the above reaction mixture at 0° C. The DIPEA (0.96 mL) was dropwiseadded to the above reaction mixture at 0° C. and the reaction mixturewas stirred for 1 hr at 0° C. to 5° C. Finally the reaction mixture wasallowed to warm to room temperature. The reaction mixture was stirredfor another 4 hrs at room temperature. Completion of reaction wasmonitored by TLC using Hexane:Ethyl acetate (7:3). The solidprecipitated out was filtered off and washed with 1-butanol (2 mL).Solid was dried under reduced pressure at 40° C. for 1 hr. ¹H-NMR(DMSO-d6, 400 MHz) δ 1.48 (S, 9H), 7.02 (m, 1H), 7.26 (m, 2H), 7.58 (S,1H), 8.57 (S, 1H), 9.48 (S, 1H), 9.55 (S, 1H).

Step 2:

To the above crude (3.1 g) in DCM (25 mL) was added TFA (12.4 mL) slowlyat 0° C. The reaction mixture was allowed to warm to room temperature.Reaction mixture was stirred for another 10 min at room temperature. Thecrude mixture was concentrated under reduced pressure.

Step 3:

The concentrated crude mixture was dissolved in DCM (25 mL) and cooledto −30° C. To the reaction mixture was slowly added acryloyl chloride(0.76 g) at −30° C. The reaction mass was warmed to room temperaturestirred at room temperature for 1.0 hr. The reaction was monitored onTLC using Hexane:Ethyl acetate (7:3) as mobile phase. Reaction gotcompleted after 1 hr. ¹H-NMR (DMSO-d6, 400 MHz) δ 5.76 (dd, J=2.0, 10.0Hz, 1 H), 6.24 (dd, J=2.0, 17.2 Hz, 1 H), 6.48 (m, 1 H), 7.14 (d, J=8.8Hz, 1 H), 7.37 (t, J=8.0 Hz, 1 H), 7.94 (S, 1 H), 8.59 (S, 1 H), 9.60(S, 1 H), 10.26 (S, 1 H).

Step 4:

To obtain the example 1, a mixture of intermediate 1 of Scheme 2 (16 mg)and intermediate 2 of Scheme 1(N1-(1-(2-fluoroethyl)azetidin-3-yl)-3-methoxybenzene-1,4-diamine, 20mg) in dioxane (1.0 mL) with catalytic trifluoroacetic acid was stirredovernight at 50° C. The crude was concentrated under reduced pressureand purified using HPLC (TFA modifier) to give the title compound (10mg) as a TFA salt. ¹H-NMR (DMSO-d6, 400 MHz) δ 10.13 (s, 1 H), 8.53 (br,1 H), 8.24 (s, 1 H), 8.07 (s, 1 H), 7.74 (br, 1 H), 7.24 (m, 3 H), 6.45(dd, J=10.4, 17.2 Hz, 1 H), 6.26 (dd, J=2.0, 17.2 Hz, 1 H), 6.18 (s, 1H), 5.92 (br, 1 H), 5.76 (dd, J=2.0, 10.4 Hz, 1 H), 4.48 (t, J=4.8 Hz, 1H), 4.36 (t, J=4.8 Hz, 1 H), 3.93 (br, 1 H), 3.69 (s, 3 H), 2.6-2.9 (m,4 H); calculated mass for C₂₆H₂₇F₄N₇O₂: 545.2, found: 546.2 (M+H⁺).

Example 2 Compound I-2N-(3-(2-(4-(1-acetylazetidin-3-ylamino)-2-methoxyphenylamino)-5-(trifluoromethyl)pyrimidin-4-ylamino)phenyl)acrylamide)

As shown in Scheme 2, a mixture of intermediate 1 of Scheme 2 (16 mg)and intermediate 3 of Scheme 1(1-(3-(4-amino-3-methoxyphenylamino)azetidin-1-yl)ethanone, 20 mg) indioxane (1.0 mL) with catalytic trifluoroacetic acid was stirredovernight at 50° C. The crude was concentrated under reduced pressureand purified using HPLC (TFA modifier) to give the title compound (12mg) as a TFA salt. ¹H-NMR (DMSO-d6, 400 MHz) δ 10.6 (s, 1 H), 10.15 (s,1 H), 9.45 (s, 1 H), 8.62 (br, 1H), 6.96-7.85 (m, 4 H), 6.55 (dd, J=2,10 Hz, 1 H), 6.23 (d, J=10 Hz, 1 H), 5.74 (d, J=10 Hz, 1 H), 3.15-3.85(m, 6 H), 1.77 (s, 3 H); calculated mass for C₂₆H₂₆F₃N₇O₃: 541.2, found:542.2 (M+H⁺).

Example 3 Compound I-35, tert-butyl3-(4-(4-(3-acrylamidophenylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3-methoxyphenylamino)azetidine-1-carboxylate)

As shown in Scheme 2, a mixture of intermediate 1 of Scheme 2 (16 mg)and intermediate 1 of Scheme 1 (tert-butyl3-(4-amino-3-methoxyphenylamino)azetidine-1-carboxylate, 20 mg) indioxane (1.0 mL) with catalytic trifluoroacetic acid was stirredovernight at 50° C. The crude was concentrated under reduced pressureand purified using HPLC (TFA modifier) to give the title compound (10mg) as a TFA salt. ¹H-NMR (DMSO-d6, 400 MHz) δ 10.1 (s, 1 H), 8.4 (br, 1H), 8.20 (s, 1 H), 8.05 (s, 1 H), 7.62 (br, 1 H), 7.43 (br, 1 H),7.23-7.26 (m, 2 H), 6.25 (dd, J=4, 10 Hz, 1 H), 6.10-6.14 (m, 1 H), 5.72(d, J=10 Hz, 1 H). 5.72 (d, J=10 Hz, 1 H), 3.67-4.03 (m, 5 H), 3.52-3.55(m, 2 H), 1.35 (s, 9 H); calculated mass for C₂₉H₃₂F₃N₇O₄: 599.3, found:600.3 (M+H⁺).

Example 4 Compound I-39,N-(3-(5-chloro-2-(4-(isopropylamino)-2-methoxyphenylamino)pyrimidin-4-yloxy)phenyl)acrylamide)

Step 1:

In a 25 mL 3-neck RBF previously equipped with a magnetic stirrer,thermo pocket and CaCl₂ guard tube, N-Boc-3-aminophenol (0.96 g) and1-butanol (9.0 mL) were charged. Reaction mixture was cooled to 0° C.2,4,5-Trichloropyrimidine (1.0 g) was added dropwise to the abovereaction mixture at 0° C. The DIPEA (0.96 mL) was dropwise added to theabove reaction mixture at 0° C. and the reaction mixture was stirred for1 hr at room temperature. Reaction mixture was stirred for 4 hrs at 50°C. Completion of reaction was monitored by TLC using Hexane:Ethylacetate (7:3). The solid precipitated out was filtered off and washedwith 1-butanol (2 mL). Solid was dried under reduced pressure at 40° C.for 1 hr. ¹H-NMR (DMSO-d6, 400 MHz) δ 1.47 (S, 9 H), 6.60 (S, 1 H), 6.86(d, J=8.4 Hz, 1 H), 7.13 (t, J=7.6 Hz, 1 H), 7.36 (m, 1 H), 7.50 (S, 1H), 8.40 (S, 1 H).

Step 2:

To the above crude (3.1 g) in DCM (25 mL) was added TFA (12.4 mL) slowlyat 0° C. The reaction mixture was allowed to warm to room temperature.Reaction mixture was stirred for another 10 min at room temperature. Thecrude was concentrated under reduced pressure.

Step 3:

The concentrated crude was dissolved in DCM (25 mL) and cooled to −30°C. To the reaction mixture was slowly added acryloyl chloride (0.76 g)at −30° C. The reaction mass was warmed to room temperature stirred atroom temperature for 1.0 hr. The reaction was monitored on TLC usingHexane:Ethyl acetate (7:3) as mobile phase. Reaction got completed after1 hr. ¹H-NMR (DMSO-d6, 400 MHz) δ 5.78 (dd, J=2.0, 10.0 Hz, 1 H), 6.25(dd, J=2.0, 17.2 Hz, 1 H), 6.40 (m, 1 H), 7.02 (d, 1 H), 7.50 (m, 2 H),7.71 (S, 1 H), 8.40 (S, 1 H), 10.35 (S, 1 H).

Step 4:

To obtain the title compound, a mixture of intermediate 1 of Scheme 3(16 mg) and N1-isopropyl-3-methoxybenzene-1,4-diamine (15 mg, preparedfrom WO 2006/021548) in dioxane (1.0 mL) with catalytic trifluoroaceticacid was stirred overnight at 100° C. The crude was concentrated underreduced pressure and purified using HPLC (TFA modifier) to give thetitle compound (6 mg) as a TFA salt. ¹H-NMR (DMSO-d6, 400 MHz) δ 10.32(s, 1 H), 8.30 (s, 1 H), 8.05 (s, 1 H), 7.62 (s, 1 H), 7.54 (d, J=8.0Hz, 1 H), 7.39 (t, J=8.4 Hz, 1 H), 7.04 (d, J=8.8 Hz, 1 H), 6.96 (d,J=8.0 Hz, 1 H), 6.43 (dd, J=10.0, 16.8 Hz, 1 H), 6.27 (dd, J=1.6, 16.8Hz, 1 H), 6.18 (s, 1 H), 5.87 (m, 1 H), 5.78 (dd, J=1.6, 10.0 Hz, 1 H),5.20 (d, J=8.0 Hz, 1 H), 3.65 (s, 3 H), 1.09 (d, J=6.4 Hz, 6 H);calculated mass for C₂₃H₂₄ClN₅O₃: 453.2, found: 454.0 (M+H⁺).

Example 5 Compound I-5,N-(3-(2-(4-(isopropylamino)-2-methoxyphenylamino)-5-(trifluoromethyl)pyrimidin-4-ylamino)phenyl)acrylamide)

As shown in Scheme 2, a mixture of intermediate 1 of Scheme 2 (16 mg)and N1-isopropyl-3-methoxybenzene-1,4-diamine (15 mg, prepared from WO2006/021548) in dioxane (1.0 mL) with catalytic trifluoroacetic acid wasstirred overnight at 50° C. The crude was concentrated under reducedpressure and purified using HPLC (TFA modifier) to give the titlecompound (8 mg) as a TFA salt. ¹H-NMR (DMSO-d6, 400 MHz) δ 10.12 (s, 1H), 8.49 (br, 1 H), 8.24 (s, 1 H), 8.06 (s, 1 H), 7.71 (br, 1 H), 7.47(d, J=6.4 Hz, 1 H), 7.23 (m, 3 H), 6.44 (dd, J=10.0, 16.8 Hz, 1 H), 6.23(m, 2 H), 5.91 (m, 1 H), 5.75 (dd, J=1.6, 10.0 Hz, 1 H), 5.21 (m, 1 H),3.68 (s, 3 H), 1.10 (d, J=6.0 Hz, 1 H); calculated mass forC₂₄H₂₅F₃N₆O₂: 486.2, found: 486.9 (M+H⁺).

Biological Examples

Described below are assays used to measure the biological activity ofprovided compounds as selective inhibitors of mutant EGFR as compared toWT EGFR (and other protein kinases).

Example 6 Omnia Assay Protocol for Potency Assessment Against EGFR (WT)and EGFR (T790M/L858R) Active Enzymes

Below describes the biochemical assay protocol using EGFR-WT andEGFR-T790M/L858R.

The mechanics of the assay platform are best described by the vendor(Invitrogen, Carlsbad, Calif.) on their website at the following URL:www.invitrogen.com/content.cfm?pageid=11338 orwww.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Drug-Discovery/Target-and-Lead-Identification-and-Validation/KinaseBiology/KB-Misc/Biochemical-Assays/Omnia-Kinase-Assays.html.

Briefly, 10× stocks of EGFR-WT (PV3872) from Invitrogen andEGFR-T790M/L858R (40350) from BPS Bioscience, San Diego, Calif., 1.13×ATP (AS001A) and appropriate Tyr-Sox conjugated peptide substrates(KCZ1001) were prepared in 1× kinase reaction buffer consisting of 20 mMTris, pH 7.5, 5 mM MgCl₂, 1 mM EGTA, 5 mM β-glycerophosphate, 5%glycerol (10× stock, KB002A) and 0.2 mM DTT (DS001A). 5 μL of eachenzyme were pre-incubated in a Corning (#3574) 384-well, white,non-binding surface microtiter plate (Corning, N.Y.) for 30 min. at 25°C. with a 0.5 μL volume of 50% DMSO and serially diluted compoundsprepared in 50% DMSO. Kinase reactions were started with the addition of45 μL of the ATP/Tyr-Sox peptide substrate mix and monitored every 71seconds for 60 minutes at λ_(ex)360/λ_(em)485 in a Synergy⁴ plate readerfrom BioTek (Winooski, Vt.). At the conclusion of each assay, progresscurves from each well were examined for linear reaction kinetics and fitstatistics (R², 95% confidence interval, absolute sum of squares).Initial velocity (0 minutes to ˜30 minutes) from each reaction wasdetermined from the slope of a plot of relative fluorescence units vstime (minutes) and then plotted against inhibitor concentration toestimate IC₅₀ from log [Inhibitor] vs Response, Variable Slope model inGraphPad Prism from GraphPad Software (San Diego, Calif.).

[EGFR-WT]=5 nM, [ATP]=15 uM, [Y12-Sox]=5 uM (ATP K_(Mapp)˜12 uM); and[EGFR-T790M/L858R]=2.5 nM, [ATP]=20 uM, [Y12-Sox]=5 uM (ATP K_(Mapp) ˜20uM).

Table 2 shows the activity of selected compounds of this invention inthe EGFR inhibition assay described above. Table 2 shows mutant EGFRdata as compared to WT EGFR and provides the selectivity ratio of WT tomutant for each test compound. The compound numbers correspond to thecompound numbers in Table 1.

TABLE 2 EGFR (Mutant and Wild Type) Biochemical Inhibition Data Com-EGFR pound WT EGFR (T790M/L858R) Ratio # IC₅₀ (nM) IC₅₀ (nM) WT/mutantI-1 1-10 <1 >1 I-2 1-10 <1 >15 I-3 100-300  1-10 >50 I-4 30-100 1-10 >25I-5 10-30  <1 >20

Example 7

Cell Culture and Antibodies

A431 human epidermoid carcinoma, H1975 human NSCLC and HCC827 humanNSCLC adenocarcinoma cells were obtained from the American Type CultureCenter (Manassas, Va.). A431 cells were grown in DMEM (Invitrogen,Carlsbad, Calif.) supplemented with 10% FBS (HyClone, South Logan, Utah)and 1% Penicillin-Streptomycin (P/S, Lonza, Walkersville, Md.). H1975and HCC827 cells were grown in complete RPMI 1640 (Invitrogen)supplemented with 10% FBS and 1% P/S. All cells were maintained andpropagated as monolayer cultures at 37° C. in a humidified 5% CO₂incubator.

All primary antibodies were obtained from Cell Signaling (Danvers,Mass.) and used at 1:1000. Secondary antibodies were used at 1:10,000.Goat anti-mouse IgG IRDye 800CW antibody was obtained from LiCorBiosciences (Lincoln, Nebr.) and goat anti-rabbit IgG Alexa Fluor 680was obtained from Invitrogen.

Immunoblotting

Cells were grown in 12-well plates (Corning, Coring, N.Y.) to 90%confluence and then incubated in low-serum (0.1% FBS) media for 16-18hr. Cells were then treated with 5, 1.25, 0.31, 0.078, 0.020 or 0.005 μMtest compound in low-serum (0.1% FBS) media for 1 hr. A431 cells werethen stimulated with 50 ng/ml EGF (Peprotech, Rocky Hill, N.J.) for 15min. After treatment, cell monolayers were washed with cold PBS(Invitrogen) and immediately lysed by scrapping into 60 uL cold CellExtraction Buffer (Invitrogen) supplemented with Complete Proteaseinhibitors (Roche, Indianapolis, Ind.) and PhosphoSTOP (Roche)phosphatase inhibitors.

Lysate protein concentrations were determined by BCA Assay (Pierce,Rockford, Ill.) and 50 ug of each lysate was separated by 4-12% gradientSDS-PAGE (Invitrogen), transferred to nitrocellulose membrane (Biorad,Hercules, Calif.) and probed with specific antibodies. Phospho-proteinsignals were quantitated using Odyssey Infrared Imaging (Li-CorBiosciences).

To assess phosphor-EGFR signaling, blots were probed with rabbitanti-Phospho-EGFR (Y1068) and mouse total anti-EGFR antibodies.Phospho-EGFR signal was normalized to total EGFR expression for eachsample. Results are indicated as % DMSO control. Normalized data wasfitted using a sigmoidal curve analysis program (Graph Pad Prism version5) with variable Hill slope to determine the EC₅₀ values.

Table 3 shows mutant EGFR data in H1975 (double mutation L858R/T790M)and HCC827 (delE746-A750 mutation) cells as compared to WT EGFR (A431cells). The compound numbers recited in Table 3 correspond to thecompound numbers in Table 1.

TABLE 3 EGFR (Mutant and Wild Type) Signaling (1 hr) Com- WT EGFR H1975HCC827 pound EC₅₀ EC₅₀ Ratio EC₅₀ Ratio # (nM) (nM) WT/mutant (nM)WT/mutant I-1 >1000  10-100 >35 100-500 >10 I-2 — — — — — I-3 >1000100-500 >10 — — I-4 — — — — — I-5 — — — — —

Example 8

Cell Proliferation

Cells were plated in Growth Media supplemented with 5% FBS and 1% P/S ata density of 3,000 cells per well in 96 well tissue culture plates(Corning). Cells were allowed to settle down for 4 hr and then treatedwith 5, 1.25, 0.31, 0.078, 0.020 or 0.005 μM test compound for 72 hr.Cell viability was determined by CellTiter Glo (Promega, Madison, Wis.)and results were converted to cell numbers using a standard curve.Growth inhibition (GI50) values were determined by Graph Pad Prism.

The result of this experiment is depicted in Table 4, where it showsmutant selective inhibition in H1975 (double mutation L858R/T790M) andHCC827 (delE746-A750 deletion mutation) cells but not in WT-EGFR A431cells.

TABLE 4 EGFR (Mutant and Wild Type) Cell Proliferation Com- WT EGFRH1975 HCC827 pound GI₅₀ GI₅₀ Ratio GI₅₀ Ratio # (nM) (nM) WT/mutant (nM)WT/mutant I-1 500-1000  10-100 >10  10-100 >10 I-2 >1000 100-500 >5100-500 >15 I-3 >1000 100-500 >5  10-100 >20 I-4 >1000  500-1000 >1100-500 >35 I-5 500-1000 100-500 >1 100-500 >5

Example 9

Washout Experiment in H1975 Cells Containing EGFR Deletion/T790MMutation

Cells are plated in Growth Media supplemented with 10% FBS and 1% P/S ata density of 2.0×10.5 cells per well in 12 well tissue culture plates.Cells are allowed to settle down for 4 hrs and then maintained inlow-serum (0.1% FBS) media overnight.

The following morning the media is removed and the cells are treatedwith 500 nM test compound in low-serum media for 1 hr. The cells arewashed free of compound 2× with PBS (Invitrogen). One set of cells areimmediately lysed as indicated above as the 0 hr time point. Theremaining cells are incubated with complete RPMI-1640 growth media (10%FBS) for 1, 3, 6 and 24 hr. For the first 1 hr, cells are washed 2× withPBS every 30 min. DMSO (0.5%) controls are collected at the 0, 3, 6 and24 hr time points.

Example 10

Mass Spectrometry for Mutant EGFR

Intact EGFR T790M/L858R (BPS Bioscience San Diego, Calif.) is incubatedfor 60 min at 37° C. with a 10-fold excess of test compound. 3 μLaliquots of the samples are diluted with 15 μL of 0.2% TFA prior toC4-Ziptipping directly onto the MALDI target plate using sinapinic acidas the desorption matrix (10 mg/mL in 0.1% TFA:acetonitrile, 50:50).

Example 11

H11975 Tumor In Vivo Study

Female nu/nu mice were implanted with 1×10⁷ H1975 tumor cells in 50%Matrigel subcutaneously (0.2 ml injection volume) in the flank. Tumormeasurements were recorded three times per week. Tumors were pairmatched when they reached an average size of 100-150 mg. Group size was10 mice. Test compound was administered intraperitoneal, 25 mg/kg or 50mg/kg daily for 21 days. % Tumor inhibition was determined at 15 daysand 18 days. Tumor volume was followed until tumors reached 1500 mm³ or60 days. Tumor inhibition value for I-1 was >80% for 25 mg/kg at both 15and 18 days, and >90% for 50 mg/kg at both 15 and 18 days.

While a number of embodiments of this invention are described herein, itis apparent that the basic examples may be altered to provide otherembodiments that utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments that have been represented by way of example.

We claim:
 1. A method of treating a proliferative disorder comprisingadministering to a patient in need thereof a compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein: R¹ is —CF₃ or—Cl; W is —O— or —NH—; R² is —R or

R³ is —R, —C(O)R, or —C(O)OR; and each R is independently C₁₋₄ alkyl orC₁₋₄ fluoroalkyl; and wherein the proliferative disorder is a cancer. 2.The method according to claim 1, wherein R¹ is —CF₃.
 3. The methodaccording to claim 1, wherein R¹ is —Cl.
 4. The method according toclaim 1, wherein R² is


5. A method of treating a proliferative disorder comprisingadministering to a patient in need thereof a compound of formula II:

or a pharmaceutically acceptable salt thereof, wherein: R³ is —R,—C(O)R, or —C(O)OR; and each R is independently C₁₋₄ alkyl or C₁₋₄fluoroalkyl; and wherein the proliferative disorder is a cancer.
 6. Themethod according to claim 1, wherein the compound is selected from:

or a pharmaceutically acceptable salt thereof.
 7. The method of claim 1,wherein the cancer is selected from breast cancer, glioblastoma, lungcancer, cancer of the head and neck, colorectal cancer, and bladdercancer.
 8. The method of claim 7, wherein the cancer is a lung cancer.9. The method of claim 8, wherein the lung cancer is non-small cell lungcancer.
 10. The method of claim 5, wherein the cancer is selected frombreast cancer, glioblastoma, lung cancer, cancer of the head and neck,colorectal cancer, and bladder cancer.
 11. The method of claim 10,wherein the cancer is a lung cancer.
 12. The method of claim 11, whereinthe lung cancer is non-small cell lung cancer.
 13. The method accordingto claim 6, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 14. The method of claim6, wherein the cancer is selected from breast cancer, glioblastoma, lungcancer, cancer of the head and neck, colorectal cancer, and bladdercancer.
 15. The method of claim 14, wherein the cancer is a lung cancer.16. The method of claim 15, wherein the lung cancer is non-small celllung cancer.